Large-scale purification of antisense oligonucleotides by high-performance membrane adsorber chromatography

Very high Aux ion-exchange membranes were utilized for a novel purification of antisense oligonucleotides (20-mer). Strong anion-exchange membranes were produced by attaching polymeric ligands onto a microporous cellulosic matrix. The oligonucleotides purified were therapeutic single-stranded phosphorothioates deoxyribonucleotides. Although small-scale membrane devices (15 cm(2)) had similar resolution to traditional chromatographic columns; their throughputs were superior. Greater than a 1300-fold scale-up produced very similar purity and yields of the phosphorothionate product. Scale-up experiments were conducted with a 2 m(2) surface area membrane module. These modules were easily capable of very high throughputs of 0.5 to 2 1/min. High purity and yields were achieved by both step and linear gradient elution. (C) 2000 Elsevier Science B.V. All rights reserved.